Protective effects of adipose-derived multipotent mesenchymal stromal cells of mice on periventricular leukomalacia model in vitro

Home/2017, Vol. 5, No. 1/Protective effects of adipose-derived multipotent mesenchymal stromal cells of mice on periventricular leukomalacia model in vitro

Protective effects of adipose-derived multipotent mesenchymal stromal cells of mice on periventricular leukomalacia model in vitro

Tsupykov O. M.1,2, Lushnikova I. V.1, Ustymenko A. M.2, Kyryk V. M.2, Nikandrova Y. A.1, Patseva M. A.1, Yatsenko K. V.1, Butenko G. M.2, Skibo G. G.1,2
1Bogomoletz Institute of Physiology NAS of Ukraine, Kyiv, Ukraine
2State Institute of Genetic and Regenerative Medicine NAMS of Ukraine, Kyiv, Ukraine


Periventricular leukomalacia (PVL) is a form of white matter lesions of the brain that results from hypoxic-ischemic injury and/or inflammation of nervous tissue, and is one of the causes of cerebral palsy. On PVL models in vivo, we have demonstrated neuroprotective effect of transplantation of adipose-derived multipotent mesenchymal stromal cells (MMSCs). However, the mechanisms, which realize neuroprotective effect of transplanted MMSCs, remain unexplored.
The aim was to assess the influence of adipose-derived MMSCs on cultured mouse brain slices at their contact co-culturing on PVL models in vitro.
Methods. Periventricular leukomalacia in vitro was modelled by a 30-minute oxygen-glucose deprivation (OGD) of mouse brain slices, followed by the addition of 100 ng/ml LPS in culture medium. For co-cultivation we used adipose-derived MMSCs obtained from mice FVB-Cg-Tg (GFPU) 5Nagy/J, transgenic for green fluorescent protein (GFP). The viability of cultured sections cells was evaluated by analysing the level of lactate dehydrogenase (LDH) in the culture medium. Probable MMSCs differentiation into neurons and glial cells was studied using immunohistochemical staining of slices using specific antibodies to neurons and oligodendrocytes (NeuN and Oligodendrocytes, respectively).
Results. Modelling of PVL in vitro on organotypic culture of brain slices led to a significant increase in level of cytosolic enzyme LDH in the culture medium. Co-cultivation of slices with MMSCs at PVL reduced the amount of this enzyme. Furthermore, it is shown that under conditions of PVL in vitro, MMSCs are able to differentiate into cells of nervous tissue.
Conclusions. Adipose-derived MMSCs have protective effect when they are co-cultivated with the mice brain slices on PVL model in vitro.

Keywords: periventricular leukomalacia; lipopolysaccharide; brain organotypic tissue culture; multipotent mesenchymal stromal cells

Full Text PDF

1. Ahn SY, Chang YS, Park WS. Mesenchymal stem cells transplantation for neuroprotection in preterm infants with severe intraventricular hemorrhage. Korean J Pediatr. 2014; 57(6): 251-56.
PMid:25076969 PMCid:PMC4115065
2. Allen M, Millett P, Dawes E, et al. Lactate dehydrogenase activity as a rapid and sensitive test for the quantification of cell numbers in vitro. Clin Mater. 1994; 16(4): 189-94.
3. Blumenthal I. Periventricular leukomalacia: a review. Eur J Pediatr. 2004; 163(8): 435-42.
4. Chernykh ER, Kafanova MY, Shevela EY, et al. Clinical experience with autologous M2 macrophages in children with severe cerebral palsy. Cell Transplant. 2014; 23(1): 97-104.
5. Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapу. 2006; 8(4): 315-17.
6. Gano D, Andersen SK, Partridge JC, et al. Diminished white matter injury over time in a cohort of premature newborns. J Pediatr. 2015; 166(1): 39–43. doi: 10.1016/j.jpeds.2014.09.009
7. Isik S, Zaim M, Yildiz MT, et al. DNA topoisomerase IIβ as a molecular switch in neural differentiation of mesenchymal stem cells. Ann Hematol. 2015; 94(2): 7-18.
8. Kan I, Melamed E, Offen D. Autotransplantation of bone marrowderived stem cells as a therapy for neurodegenerative diseases. Handb Exp Pharmacol. 2007; 180: 219-42.
9. Khwaja O, Volpe JJ. Pathogenesis of cerebral white matter injury of prematurity. Arch Dis Child Fetal Neonatal Ed. 2008; 93(2): 153-161. doi: 10.1136/adc.2006.108837
10. Kim JM, Lee ST, Chu K, et al. Systemic transplantation of human adipose stem cells attenuated cerebral inflammation and degeneration in a hemorrhagic stroke model. Brain Res. 2007; 1183: 43-45.
11. Knuesel I, Chicha L, Britschgi M, et al. Maternal immune activation and abnormal brain development across CNS disorders. Nat Rev Neurol. 2014; 10(11): 643-60.
13. Lee JA, Kim BI, Jo CH, et al. Mesenchymal stem-cell transplantation for hypoxic-ischemic brain injury in neonatal rat model. Pediatric Research. 2010; 67(1): 42-46.
14. Lindvall O, Kokaia Z. Stem cells in human neurodegenerative disorders – time for clinical translation? J Clin Invest. 2010; 120(1): 29-40.
PMid:20051634 PMCid:PMC2798697
15. Linero I, Chaparro O. Paracrine effect of mesenchymal stem cells derived from human adipose tissue in bone regeneration. PLoS One. 2014; 9(9): 107001.
PMid:25198551 PMCid:PMC4157844
16. Rasmussen JG, Frøbert O, Pilgaard L, et al. Prolonged hypoxic culture and trypsinization increase the pro-angiogenic potential of human adipose tissue-derived stem cells. Cytotherapy. 2011; 13(3): 318-28.
17. Ryu HH, Lim JH, Byeon YE, et al. Functional recovery and neural differentiation after transplantation of allogenic adipose-derived stem cells in a canine model of acute spinal cord injury. J Vet Sci. 2009; 10(4): 273-84.
PMid:19934591 PMCid:PMC2807262
18. Sadan O, Melamed E, Offen D. Bone-marrow-derived mesenchymal stem cell therapy for neurodegenerative diseases. Expert Opin Biol Ther. 2009; 9(12): 1487-497.
19. Tsai HL, Deng WP, Lai WF, et al. Wnts enhance neurotrophin-induced neuronal differentiation in adult bone-marrow-derived mesenchymal stem cells via canonical and noncanonical signaling pathways. PLoS One. 2014; 9(8): e104937.
PMid:25170755 PMCid:PMC4149376
20. Tsupykov OM, Kyryk VM, Ustymenko AM, et al. Effect of transplantation of adipose-derived multipotent mesenchymal stromal cells on the nervous tissue and behavioral responses in a mouse model of periventricular leukomalacia. Cell and Organ Transplantology. 2015; 3(1): 68-73.
21. Tsupykov OM, Lushnikova IV, Nikandrova YA, et al.A novel model of periventricular leukomalacia on mouse organotypic brain slice culture. Cell and Organ Transplantology. 2016; 4(2): 188-193. doi:10.22494/cot.v4i2.60
22. Velthoven CTJ, Kavelaars A, Heijnen CJ. Mesenchymal stem cells as a treatment for neonatal ischemic brain damage. Pediatric Research. 2012; 71(4): 474-81.
23. Volpe JJ. Brain injury in premature infants: a complex amalgam of destructive and developmental disturbances. Lancet Neurol. 2009; 8(1): 110-124. doi: 10.1016/S1474-4422(08)70294-1
24. Wang LW, Lin YC, Wang ST, et al. Hypoxic/ischemic and infectious events have cumulative effects on the risk of cerebral palsy in very-low-birth-weight preterm infants. Neonatology. 2014; 106(3): 209-215.

Tsupykov OM, Lushnikova IV, Ustymenko AM, Kyryk VM, Nikandrova YA, Patseva MA, Yatsenko KV, Butenko GM, Skibo GG. Protective effects of adipose-derived multipotent mesenchymal stromal cells of mice on periventricular leukomalacia model in vitro. Cell and Organ Transplantology. 2017; 5(1):28-32. doi:10.22494/cot.v5i1.66


Creative Commons License
Is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.